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Cancer Genomics Proteomics. Author manuscript; available in PMC 2015 April 24. Published in final edited form as: Cancer Genomics Proteomics. 2014 ; 11(4): 175–194.

The Metastasis Suppressor NME1 Regulates Expression of Genes Linked to Metastasis and Patient Outcome in Melanoma and Breast Carcinoma JOSEPH R. MCCORKLE1, MARY K. LEONARD2, SUSAN D. KRANER4, ERIC M. BLALOCK4, MA DEQIN5, STEPHEN G. ZIMMER, and DAVID M. KAETZEL2,3,*

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1Department

of Pharmaceutical Sciences, St. Jude Children’s Research Hospital, Memphis, TN,

USA 2Department 3Marlene

of Biochemistry and Molecular Biology, University of Maryland, Baltimore, MD USA

and Stewart Greenebaum Cancer Center, University of Maryland, Baltimore, MD USA

4Department

of Molecular and Biomedical Pharmacology, College of Medicine, University of Kentucky, Lexington, KY, USA 5Department

of Pathology, University of Iowa Hospitals and Clinics, University of Iowa Carver College of Medicine, Iowa City, IA, USA

Abstract Author Manuscript Author Manuscript

NME1 is a well-documented metastasis suppressor gene, with suppressor activity demonstrated across a wide spectrum of human cancers including melanoma and carcinomas of the breast, stomach and thyroid. A primary aim of the current study was to identify profiles of genes whose expression is regulated by NME1 in cell lines of melanoma and thyroid carcinoma origin. Impact of NME1 was determined by forcing its expression transiently in cell lines using a novel Ad5based adenoviral vector (Ad5-NME1), followed 48 h later by analysis of RNA expression profiles using the U133A microarray chip. Robust NME1 expression was achieved following infection with the Ad5-NME1 adenovirus in the human metastasis-derived cell lines WM1158 (melanoma) and WRO82 (follicular thyroid carcinoma), resulting in wide-ranging effects on gene expression in both settings. A substantial proportion of the NME1-regulated genes identified in the analyses were of clear potential relevance to metastasis, such as matrix metalloproteinase-1 (MMP1), angiopoeitin-2 (ANGPT2), SERPINB9 and colony stimulating factor receptor-2B (CSFR2B). Nine genes were identified (false discovery rate ≥0.1) that were regulated by NME1 in both the WM1158 and WRO82 cell lines, each possessing one of more such metastasis-relevant activities as stress fiber formation and focal adhesion (PPM1E, ZYX, PFN1), chemotaxis (CCR1) epithelialmesenchymal signaling (WNT1), differentiation and morphogenesis (TBX4, ZFP36L2), and G protein modulation (GPR52 and PFN1). In addition, a number of the NME1-regulated genes were shown to be of prognostic value for distant disease-free survival and overall survival in melanoma and breast cancer. The combined expression of three NME1-regulated genes CSFR2B, MSF4A1 and SERPINB9 provided a strongly synergistic correlation with distant disease-free survival in the

Correspondence to: David M. Kaetzel, PhD, Department of Biochemistry and Molecular Biology, University of Maryland, Baltimore, Maryland 21201, USA, Tel: +410-706-5080; Fax: +410-706-8297; [email protected].

MCCORKLE et al.

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basal subtype of breast cancer (p90% of infected cells in both lines, consistent with attainment of NME1 overexpression in a similar proportion of the cells (Figure 1C). Profiling of NME1-regulated gene expression in WM1158 and WRO82 cells

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For RNA expression analysis, WM1158 and WRO82 cells were infected with the control adenovirus Ad5-clig, Ad-NME1, or were left uninfected, followed 48h later by harvesting of total cellular RNA (see Materials and Methods). RNA samples were analyzed using the U133A human microarray chip (Affymetrix), which analyzes expression for approximately 18,400 transcripts and variants. Genes were identified as NME1-regulated based on the following criteria. Signal intensities for the three experimental groups were compared pairwise using Welch two sample t-tests. Resulting p-values were adjusted for multiple comparisons using the correction method of Benjamini and Hochberg (26) and are listed as FDR p-values herein. Probe sets showing significant differences (unadjusted p

The metastasis suppressor NME1 regulates expression of genes linked to metastasis and patient outcome in melanoma and breast carcinoma.

NME1 is a well-documented metastasis suppressor gene, with suppressor activity demonstrated across a wide spectrum of human cancers including melanoma...
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